The present invention relates to antimicrobial cephalosporins.
In one aspect the present invention provides a compound of formula 
wherein
R1 denotes hydrogen, acyl, carboxyl, or alkyl;
R2 and R3 are the same or different and independently of each other denote hydrogen, cycloalkyl, alkyl, alkenyl or alkinyl;
R4 denotes hydrogen or a group of formula 
wherein R6 denotes amino, hydrazino, aminoalkylamino, alkoxy, aryl, cycloalkyl, aryloxy, heterocyclyl, alkyl, alkenyl, alkinyl;
R5 denotes hydrogen or an ester moiety;
W denotes CH or N;
V denotes CH or Nxe2x80x94O; and
Z denotes O, S or NR7, wherein R7 is as defined as R2;
with the proviso that not all of R2, R3 and R4 denote hydrogen; and,
if R4 denotes hydrogen, R1 is other than H or CH3.
A compound of formula I includes a compound of formula 
wherein W and R5 are as defined above,
Rxe2x80x21 denotes hydrogen or alkyl, e.g. including unsubstituted alkyl, e.g. (C1-12)alkyl, such as lower alkyl; or substituted alkyl by e.g. halogen, carboxy; e.g. hydrogen or CH2F;
Rxe2x80x22 and Rxe2x80x23 are the same or different and independently of each other denote hydrogen; alkenyl; e.g. (C2-4)alkenyl; or alkyl;
e.g. unsubstituted or substituted by e.g. halogen, aryl; preferably aryl; including unsubstituted aryl, or aryl substituted by e.g. alkoxy, such as C(1-4)alkoxy or hydroxy; e.g.
Rxe2x80x22 denotes hydrogen, alkyl, or alkenyl; and
Rxe2x80x23 denotes hydrogen or alkyl; and
Rxe2x80x24 denotes hydrogen or a group of formula. 
wherein
Zxe2x80x2 denotes O or NRxe2x80x27, wherein Rxe2x80x27 denotes hydrogen or alkyl, e.g. lower alkyl; and
Rxe2x80x26 denotes amino, including e.g.(di)lower alkylamino; aminoalkylamino, including e.g ((di)-lower alkyl)amino-(lower)alkylamino; hydrazino; alkoxy, e.g. lower alkoxy; unsubstituted aryl or aryl substituted e.g. by (lower alkyl)carbonyloxy, lower alkoxy; cycloalkyl; a 5 to 6 membered, heterocycle containing 1 to 3 nitrogen and/or sulphur-and/or oxygen atoms, e.g. 1 to 3 nitrogen atoms such as pyrrolidinyl;
alkyl, alkenyl, alkinyl including alkyl, alkenyl, alkinyl interrupted by N, S and/or O; e.g. unsubstituted alkyl, alkenyl, alkinyl or substituted alkyl, alkenyl, alkinyl by hydroxy, aryl, hydroxyaryl, guanidino, nitroguanidino, alkoxy, aryloxy, acyloxy, carbamoyloxy, amino, alkylamino, dialkylamino, trialkylammonium, acylamino, ureido, alkoximino, oximino, imino, carboxy, oxo, halogen, nitro, a carboxylic acid derivative, a sulphonic acid derivative, or heterocyclyl;
such as alkyl, e.g. substituted alkyl, e.g. one or several-fold; by unsubstituted aryl, or substituted aryl by hydroxy, alkoxy, phenoxy; aryloxy, e.g. phenoxy; amino, including e.g. (di)lower alkylamino; hydroxy; carboxy; guanidino or nitroguanidino; or a heterocyclyl-carboximino group; with the proviso that not all of R2, R3 and R4 denote hydrogen.
If not otherwise defined herein any aliphatic group defined herein includes an aliphatic group containing up to 20, e.g. 12, such as 8 C-atoms. Acyl includes aliphatic or aromatic acyl. Lower alkyl includes (C1-4)alkyl. Aryl includes aryl containing up to 18, e.g. 12 C atoms, including e.g. phenyl, napthyl. Cycloalkyl includes (C3-8)cycloalkyl, such as (C3-6)cycloalkyl.
Heterocyclyl includes e.g. saturated or (partially) unsaturated heterocyclyl having 5 or 6 ring members and 1 to 5, e.g. 1 to 3 nitrogen and/or 1 to 3 sulphur and/or oxygen hetero atoms including, for example, condensed heterocyclyl, such as benzthiazolyl. Any group as defined may be unsubstituted or substituted, e.g. by groups which are conventional groups in xcex2-lactam chemistry. Substituted heterocyclyl includes preferably substituted heterocyclyl by amino, hydroxy, alkoxy, acyloxy, carboxy or mercapto. An ester-moiety includes alkyl, preferably C1-6alkyl, e.g. C1-4alkyl; aralkyl, for example benzyl, alkoxybenzyl, such as 4-methoxybenzyl; indanyl, phthalidyl, alkoxymethyl, e.g. methoxymethyl; (C1-6)alkanoyloxy(C1-6)alkyl, (C1-6)alkoxy-carbonyl-oxy(C1-6)alkyl, glycyloxymethyl, phenylglycyloxymethyl, (5-methyl-2-oxo-1,3-dioxolen4-yl)methyl; an ester moiety also includes ester moieties which form with the COOxe2x80x94group a physiologically hydrolysable and acceptable ester, e.g. such known to be hydrolysable ester groups in the field of cephalosporins. A compound of formula I may thus be in the form of an physiologically-hydrolysable and -acceptable ester. By physiologically-hydrolysable and -acceptable esters as used herein is meant an ester in which the COOxe2x80x94 group is esterified and which is hydrolysable under physiological conditions to yield an acid which is itself physiologically tolerable at dosages to be administered. The term is thus to be understood as defining regular pro-drug forms. An ester moiety may be preferably a group which is easily hydrolysable under physiological conditions. Such esters may be administered preferably orally. Parenteral administration may be indicated if the ester per se is an active compound or, if hydrolysis occurs in the blood. A silyl group includes a silyl protecting group, e.g. a conventional silyl protecting group, such as a trialkylsilyl group, for example the trimethylsilyl group. A leaving group includes e.g. a leaving group which is conventional in a type of reaction described; in an acylation reaction of an amine group e.g. a carboxylic acid derivative, such as a carboxylic acid halogenide, (active) ester, (mixed) anhydride) may be an appropriate acylation agent. A cation includes a cation which may form a pharmaceutically acceptable salt with a compound of formula I; e.g. a metal salt such as sodium, potassium; or an amine (ammonium) salt, such as trialkylamine, procain, dibenzylamine, benzylamine, ammonium salt.
A compound of formula I includes a compound of formula 
wherein
R5 is as defined above;
R2s and R3s independently of each other denote alkyl, e.g. C1-6alkyl, such as lower alkyl; cycloalkyl, aralkyl, e.g. ar(C1-6)alkyl, such as ar(C1-4)alkyl; aryl; alkenyl, e.g. (C2-6)alkenyl, such as (C2-4) alkenyl; or alkinyl; and R3s additionally denotes hydrogen; e.g. R2s denotes alkyl, alkenyl or aralkyl; e.g. R3s denotes hydrogen or alkyl; e.g. a compound of formula 
wherein R5 is as defined above.
A compound of formula I includes a compound of formula 
wherein R1, R5, W and V are as defined above,
R2p and R3p are the same or different and independently of each other denote hydrogen, cycloalkyl, or substituted alkyl by halogen or hydroxy,
R6p denotes amino, unsubstituted or substituted alkylamino or dialkylamino, alkoxy, aryl, cycloalkyl, aryloxy, an unsubstituted, 5- or 6-membered, saturated, partially saturated or unsaturated heterocycle which may be condensed containing 1 to 5 nitrogen and/or 1 to 3 sulphur- and/or oxygen atoms, a substituted 5- or 6-membered, saturated, partially saturated or unsaturated heterocycle which may be condensed containing 1 to 5 nitrogen and/or 1 to 3 sulphur- and/or oxygen atoms by amino, hydroxy, alkoxy, acyloxy, carboxy or mercapto, cycloalkyl or unsubstituted straight chain or branched (C1-20)alkyl, (C1-20)alkenyl or (C1-20)alkinyl, which may be interrupted by N, S and/or O, once or several times, substituted straight chain or branched (C1-20)alkyl, (C1-20)alkenyl or (C1-20)alkinyl, which may be interrupted by N, S and/or O, by hydroxy, alkoxy, aryloxy, acyloxy, carbamoyloxy, amino, alkylamino, dialkylamino, trialkylammonium, acylamino, ureido, oximino, imino, carboxy, oxo, halogen, nitro, a carboxylic acid derivative, a sulphonic acid derivative, an unsubstituted, 5- or 6-membered, saturated, partially saturated or unsaturated heterocycle containing 1 to 5 nitrogen and/or 1 to 3 sulphur- and/or oxygen atoms which may be condensed; or a substituted 5- or 6-membered, saturated, partially saturated or unsaturated heterocycle which may be condensed containing 1 to 5 nitrogen and/or 1 to 3 sulphur- and/or oxygen atoms by amino, hydroxy, alkoxy, acyloxy, carboxy or mercapto;
Zp denotes oxygen or NR7p, wherein R7p is as defined R2p.
A compound of formula I includes a compound of formula 
wherein W and R5 are as defined above,
R1p denotes hydrogen or CH2F, and
Rxe2x80x26p denotes hydrogen, (C1-20)alkyl, one or two fold substituted (C1-20)alkyl by phenyl, phenoxy, amino, hydroxyphenyl, hydroxy, carboxyl, guanidino or nitroguanidino, unsubstituted phenyl or substituted phenyl by acetoxy, pyrrolidinyl; or a compound of formula 
A compound of formulae I includes a compound of formulae Ia, Is, Ip1, Ip2, and Ip3 and may be e.g. in free form and in the form of a salt and/or in the form of a solvate. A salt includes any possible salt, e.g. an acid addition salt; such as a hydrochloride, internal salt, metal salt, quaternary salt and an amine salt of a compound of formula I. Metal salts include for example sodium, potassium, calcium, barium, zinc, aluminum salts, preferably sodium or potassium salts. Amine salts include for example trialkylamine, procaine, dibenzylamine and benzylamine salts. A salt may preferably be a pharmaceutically acceptable salt of a compound of formula I.
A solvate includes a solvate with an organic solvent and a solvate with water, such as a hydrate.
A compound of formula I may be e.g. in the form of a hydrochloride, such as a monohydrochloride, dihydrochloride, trihydrochloride, e.g. in crystalline form and/or in the form of a solvate, e.g. a hydrate.
A free form of a compound of formula I may be converted into a salt form and vice versa. A solvate form of a compound of formula I, e.g. in free form or in the form of a salt, may be converted in a non-solvate form and vice versa.
A compound of formula I includes a compound of formula I in any configuration, e.g. in any possible steroisomeric form. Mixtures of stereoisomeric forms may be separated, e.g. as conventional, e.g. by chromatography, fractioned crystallisation. E.g. the configuration of R1 in group xe2x80x94Cxe2x95x90VR1 may be syn [(Z)] and anti [(E)] and is preferably, e.g. predominantly, syn [(Z)]; e.g. containing the [(E)] form in an amount of 0 to 5%, e.g. 0 to 2%.
A compound of formula I may be in the form of a mixture of the 3(E)-form and 3-(Z)-form, or may be, e.g. predominantly, in the 3(Z)-form, e.g. according to formula 
or may be, e.g. predominantly, in the 3(E)-form, e.g. according to formula 
wherein R1 and R2 are as defined above, and wherein the configuration of the group 
attached to the nitrogen of the xe2x80x94Cxe2x95x90N group in position 3 of the ring system is, e.g. 3(E) and/or 3(Z). A compound of formula I may be, e.g. predominantly, in the 3(E)-form, e.g. containing the 3(Z)-form in an amount of 0 to 5%, e.g. 0 to 2% or predominantly in the 3(Z)-form, e.g. containing the 3(Z)-form in an amount of 0 to 5%, e.g. 0 to 2%. A compound of formulae Is and Ip1 may be, e.g. predominantly, in the 3(E)-form, e.g. containing the 3(Z)-form in an amount of 0 to 5%, e.g. 0 to 2%.
A compound of formula I may be obtained as follows:
a) Reacting a compound of formula 
wherein W, V and R1 are as defined above and wherein
xcex1) Rb denotes hydroxy and Rc and Rd together denote a bond, or
xcex2) Rd denotes hydrogen, a cation, an ester moiety or a silyl group and Rb and Rc denote the oxo group
e.g. in free form or in the form of an acid addition salt, with a compound of formula 
wherein R2, R3 and R4 are as defined above, e.g. in free form or in the form of an acid addition salt, e.g. as appropriate, e.g. as conventional
b) for the production of a compound of formula 
wherein W, V, Z, R1, R2, R3, R5 and R6 are as defined above, acylating a compound of formula 
wherein Z, R2, R3, R5 and R6 are as defined above, e.g. in free form or in the form of an acid addition salt, e.g. as appropriate, e.g. as conventional with a compound of formula 
wherein V, W and R1, are as defined above and X denotes a leaving group, e.g. in free form or in the form of an acid addition salt; e.g. as appropriate, e.g. as conventional; or reacting a compound of formula 
wherein R1, R2, R3, R5, V and W are as defined above, e.g. in free form or in the form of an acid addition salt, e.g. as appropriate, e.g. as conventional with a compound of formula 
wherein R6 and Z are as defined above and X denotes a leaving group.
Reactive groups in a compound of of formulae I, Ib, Ic, II, III, IV, V and Va may be protected by protecting groups, e.g. protecting groups which are conventional, e.g. in cephalosporin chemistry. Silyl protecting group technology in the presence of a solvent which may be inert towards silylation agents, e.g. a chlorinated hydrocarbon, such as dichloromethane; a nitrile such as acetonitrile, an ether such as tetrahydrofuran, a dipolar aprotic solvent, e.g. N,N-dimethylformamide; or a solvent system, e.g. mixtures of individual solvents, e.g. as described above; may be appropriate for the protection of reactive groups. Protecting groups may be split off, e.g. as conventional during a corresponding reaction or after termination of a corresponding reaction. A compound of formula I wherein R5 denotes hydrogen may be converted into a compound of formula I wherein R5 denotes an ester moiety or vice versa. A compound of formula I may be isolated from the reaction mixture, e.g. as conventional. A compound of formula I may be obtained in free form or in the form of a salt and/or a hydrate. A compound of formula I in free form may be converted into a compound of formula in the form of a salt and/or a hydrate and vice versa.
Process a) may be carried out as follows:
A compound of formula II may be reacted with a compound of formula III, e.g. in a solvent, e.g. in a solvent which is inert under the reaction conditions, such as water; a mixture of water with an e.g. lower, e.g. (C1-4)alcohol or dioxane; or in a dipolar aprotic solvent, e.g. dimethylformamide, dimethylsulfoxide, dimethylacetamide, if desired in mixture with an alcohol and/or water; at temperatures from xe2x88x9220 to 50xc2x0 C. The pH may be at an optimum, e.g. by addition of an organic or inorganic acid or base. A compound of formula I obtained may be isolated and/or purified, e.g. as conventional, e.g. by addition of an anti-solvent or by chromatography.
Process b) may be carried out e.g. as conventional for an acylation reaction. E.g. a compound of formula IV may be reacted with a compound of formula V; or a compound of formula Ic may be reacted with a compound of formula Va; e.g. in an appropriate solvent, such as a mixture of water and acetone or acetonitrile at appropriate temperatures, e.g. at room temperature.
Starting compounds are known or may be produced according to known, e.g. analogous methods, or e.g. according to the present examples. A part of the starting compounds according to the present invention is novel.
In another aspect the present invention provides a compound selected from
1-[(1-Methylhydrazino)iminomethyl]piperazine
1-[(1-Ethylhydrazino)iminomethyl]piperazine
1-[(1-Allylhydrazino)iminomethyl]piperazine
1-[(1-(4-Methoxybenzyl)hydrazino]iminomethyl]piperazine
1-[(1-(4-Methoxybenzyl)hydrazino]iminomethyl]piperazine
1-[(1-(3,4,5-Trimethoxybenzyl)hydrazino]iminomethyl]piperazine
1-[(1-Methylhydrazino)(methylimino)methyl]piperazine
Glycin-(4-hydrazinoiminomethyl)piperazide
1-(R)-(Amino(4-hydroxyphenyl)acetyl)4-(hydrazinoiminomethyl)piperazine
1,4-bis-(Hydrazinoiminomethyl)piperazine
1-(Hydrazinoiminomethyl)4-[ethyfimino)[3-dimethylaminopropyl)amino]methyl]piperazine, e.g. in the form of a salt, such as a hydrochloride, and/or in the form of a solvate; and, in another aspect, a compound of formula 
wherein R5 is as defined in claim 1, and Rint denotes a group 
which is formed by a bond of the terminal amine group of the hydrazino group of a compound selected from the list above and wherein the xe2x80x94Nxe2x80x94 group is substituted according to a compound selected from the list above, i.e. a hydrazino-compound listed above is bond to the ring system via the terminal amine group of the hydrazino group to the methyl group in position 3 of the ring system to form a group 
wherein the xe2x80x94Nxe2x80x94 group is substituted according to a hydrazino compound listed above.
The compounds of formulae I, hereinafter designated as xe2x80x9cactive compound(s)) of the inventionxe2x80x9d exhibits pharmacological activity and surprising low toxicity and are therefore useful as pharmaceuticals. In particular, the active compounds of the invention show antimicrobial, e.g. antibacterial, activity against aerobic and anaerobic growing bacteria, e.g. gram negative and gram positive bacteria such as Enterobacter, e.g. Enterobacter cloacae; Enterococcus, e.g. Enterococcus faecalis, Enterococcus faecium; Moraxella, e.g. Moraxella catarrhalis; Haemophilus, e.g. Haemophilus influenza; Klebsiella, e.g. Klebsiella edwardsii, Klebsiella pneumoniae; Streptococcus, e.g. Streptococcus pyogenes; Staphylococcus, e.g. Staphylococcus aureus MSSA (methicillin sensitive strains); Staphylococcus aureus MRSA (methicillin resistant strains); Escherichia, e.g. Escherichia coli; Proteus, e.g. Proteus mirabilis, Salmonella, e.g. Salmonella typhimurium, Serratia, e.g. Serratia marcescens, Pseudomonas, e.g. Pseudomonas aeruginosa; Pneumococci, e.g. Pneumococcus pneumoniae (penicillin sensitive and mult-drug resistant strains); in vitro in the Agar DilutionTest for bacteria according to National Commitee for Clinical Laboratory Standards (NCCLS) 1993,
Document-M7-A3 Vol.13, No. 25: xe2x80x9cMethods for dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically-Third Edition, Approved Standardxe2x80x9d; and
Document M11-A3 for anaerobic bacteria
in a concentration from about 0.001 to ca. 50 xcexcg/ml (MIC), e.g. using strains including Staphylococcus aureus (ATCC 29213 and ATCC 9144); Enterococcus faecalis (ATCC 29212); Haemophilus influenza (NTCC 49247 and NCTC 11931); Escherichia coli (ATCC 25922 and ATCC 35218); Klebsiella pneumoniae (NCTC 11228); Klebsiella edwardsii (NCTC 10896); Pseudomonas aeruginosa (ATCC 27853 and ATCC 25668); and in vivo in the septicaemia mouse model, in accordance to the method description Nr. 159 A-5, approved by Austrian Health Authorities (MA 58, no. 2968/95 of 12-Oct-1995), e.g. when administerd at dosages from about 0.05 to 50 mg/kg body weight, such as 0.1 to 50 mg/kg body weight (ED50 values). E.g., mice are infected with an ED 95% of Staphylococcus aureus (ATCC 4995), Streprococcus pyogenes (ATCC 29218), Escherichia coli (xcex94 12 NFI culture collection) and are treated 1, 5 and 24 hours after infection. The ED 50% values ranging from ca. 0.2 to 50 mg/kg body weight are calculated by Probit analysis of the administered dosages of compounds. Activity is determined by numbers of surviving animals per group of 8 mice per dosage until day 5 after infection.
The active compounds of the invention show an surprising overall activity spectrum. It has, for example, been determined that the MHK (xcexcg/ml) of the compound of Example 1 against, for example Enterococcus faecalis is of ca. 0.1 to 0.4; against Staphylococcus aureus (MSSA) is of ca.  less than 0.125 to 0.8; against methicillin resistant Staphyloccous aureus is of 0.8 to 6.4; against multi-drug resistant Pneumococcus is of 0.4.
The active compounds of the invention are therefore useful for the treatment of microbial, e.g. bacterial diseases.
For this indication, the appropriate dosage will, of course, vary depending upon, for example, the compound of formula I employed, the host, the mode of administration and the nature and severity of the conditions being treated. However, in general, for satisfactory results in larger mammals, for example humans, an indicated daily dosage is in the range from about 0.05 to 5 g, for example 0.1 to about 2.5 g, of an active compound of the invention conveniently administered, for example, in divided doses up to four times a day.
An active compound of the invention may be administered by any conventional route, for example orally, e.g. in the form of tablets or capsules, or parenterally in the form of injectable solutions or suspensions, e.g. in analogous manner to cefotaxime.
The compound 7-(((5-amino-1,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxyimino)acetyl)amino)-3-((imino-1-piperazinylmethyl)methylhydrazono)methyl-3-cephem4-carboxylic acid (compound of Example 1) is the preferred compound of the invention for use as an antimicrobial agent. It has, for example been determined that the MHK (xcexcg/ml) of the compound of Example 1 (tested in the form of the hydrochloride) against, for example Haemophilus influenza is ca.  less than 0.125 to 0.4 and, for example cefotaxime shows an MHK (xcexcg/ml) of ca.  less than 0.125 to 0.4. It is therefore, indicated that for the treatment of microbial diseases, e.g. bacterial diseases the preferred compounds of the invention may be administered to larger mammals, for example humans, by similar modes of administration at similar dosages than conventionally employed with cefotaxime.
A compound of formula I may be administered in the form of a pharmaceutically acceptable salt, e.g. an acid addition salt or a base addition salt or in the corresponding free form, if desired in the form of a solvate. Such a saltisolvate may exhibit the same order of activity as the free form.
The present invention also provides a pharmaceutical composition comprising a compound of formula I according to claim 1 in the form of a pharmaceutically acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent.
Such compositions may be manufactured in conventional manner. Unit dosage form may contain, for example 10 mg to about 1 g, for example 10 mg to about 700 mg, such as to about 500 mg.
As medicaments, the active ingredients according to the invention may be administered alone or in suitable medicinal forms together with inorganic or organic, pharmacologically inert excipients. For example, they are used as a constituent of capsules, or injection or instillation preparations, which contain a quantity of active compounds that is sufficient to attain an optimum blood level, that is, ca. 10 to 500 mg per capsule. For this application, the dosage to be administered depends on the compound used and the type of administration, as well as the type of treatment. With larger mammals, satisfactory results may be obtained when administering a daily dose of ca. 0.5 to 6 g. If required, this amount may be given in correspondingly smaller doses two to four times daily, or in sustained release form.
In another aspect the present invention provides a compound of formula I or a composition comprising a compound of formula I in the form of a pharmaceutically acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent for use as a pharmaceutical, e.g. as an antibiotic; and
The use of a compound of formula I, or use of a composition comprising a compound of formula I in the form of a pharmaceutically acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent as a pharmaceutical.
In a further aspect the present invention provides a method of treatment of microbial diseases, e.g. caused by bacterias selected from Pseudomonas, Enterobacter, Enterococcus, Moraxella, Haemophilus, Klebsiella, Streptococcus, Staphylococcus, Escherichia, Proteus, Salmonella, Serratia or Pneumococci, which comprises administering to a subject in need of such treatment, an effective amount of a compound of formula I; e.g. in the form of a pharmaceutical composition according to the present invention; and
A compound of formula I for use in the preparation of a medicament for the treatment of microbial diseases, for example of diseaeses caused by bacterias selected from Pseudomonas, Enterobacter, Enterococcus, Moraxella, Haemophilus, Klebsiella, Streptococcus, Staphylococcus, Escherichia, Proteus, Salmonella, Serratia or Pneumococci.